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Table 1 List of oligonucleotides used for MSP, MCA-Meth and bisulfite sequencing of RASSF1A, MGMT and BLU genes.

From: Detection of methylation in promoter sequences by melting curve analysis-based semiquantitative real time PCR

 

MSP and MCA-MSP

MCA-Meth

bisulfite sequencing

RASSF1A

UF: 5'-GAGAGTGTGTTTAGTTTTGTTTTT-3'

F: 5'-AGTTTTTGTATTTAGGTTTTTATTG-3'

F: 5'-ATAGTAAAGTTGGTTTTTAGAAATA-3'

 

UR: 5'-CCCATACTTCACTAACTTTAAACAC-3'

R: 5'-AACTCAATAAACTCAAACTCCCC-3'

R: 5'-CAACTCAATAAACTCAAACTCCCCC-3'

 

MF: 5'-GAGAGCGCTTTAGTTTCGTTTTC-3'

  
 

MR: 5'-ACCCGTACGTTCGCTAACTTTAAACG-3'

  

MGMT

UF: 5'-GAGAGATTTGTGTTTTGGGTTTAGTG-3'

F: 5'-GGTTTGGGGGTTTTTGATTAG-3'

F1: 5'-GGTATTAGGAGGGGAGAGATT-3'

 

UR: 5'-CCTTCAACCAATACAAACCAAACAA-3'

R: 5'-CCTTTTCCTATCACAAAAATAATCC-3'

R1: 5'-TATACCTTAATTTACCAAATAACCC-3'

 

MF: 5'-ATTCGCGTTTCGGGTTTAGC-3'

 

F2: 5'-GTAAATTAAGGTATAGAGTTTTAGG-3'

 

MR: 5'-CGACCGATACAAACCGAACG-3'

 

R2: 5'-AACTATCCCAACATATCC-3'

BLU

UF: 5'-TTTGTGGGTTATAGTTTGAGAAAGTG-3'

F: 5'-AAGGATTTGGAGTTTAGGAGAGATT-3'

F: 5'-AAGGATTTGGAGTTTAGGAGAGATT-3'

 

UR: 5'-AACAAATTAACCACACCTACAC-3'

R: 5'-CCAAAATCTAAAACAAAACAATTAC-3'

R: 5'-CCAAAATCTAAAACAAAACAATTAC-3'

 

MF: 5'-TTCGTGGGTTATAGTTCGAGAAAGCG-3'

  
 

MR: 5'-AACGAATTAACCGCGCCTACGC-3'

  
  1. Annealing temperatures for every set of primers:
  2. MSP: RASSF1A UF and UR: 62°C; RASSF1A MF and MR:58°C; MGMT UF and UR: 62°C; MGMT MF and UR: 62°C; BLU UF and UR: 60°C; and BLU MF and MR: 66°C.
  3. MCA-Meth: RASSF1A F and R: 58°C; BLU F and R: 56°C; and MGMT F and R: 61°C.
  4. Bisulfite sequencing: RASSF1A F and R: 61°C; MGMT F1 and R1: 58°C; MGMT F2 and R2: 58°C; and BLU F and R: 56°C.
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BMC Cancer

ISSN: 1471-2407

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